Benzo (k) fluoranthene (BkF) has adverse effects on male reproduction, but its specific mechanism of action is still unclear. This study focused on the role of RNA reading protein YTHDF2 and its mechanism in BkF induced male reproductive injury. Mouse GC-2 spermatocytes were exposed to 0, 40, 80, 160 μM BkF. It was found that BkF significantly increased the apoptosis of GC-2 cell and decreased its survival rate. BCL2 in spermatocytes decreased significantly, while the expression of P53 and BAX exhibited a notable increase. Interestingly, the expression of RNA reading protein YTHDF2 progressively rose in tandem with the escalating BkF exposure dosage. Overexpression of YTHDF2 significantly reduced the viability of cells and increased the apoptosis rate. Meanwhile, there was a substantial increase in the expression of P53 and BAX, BCL2 was significantly down-regulated. On the contrary, interfering with YTHDF2 increased cell proliferation and reduced cell apoptosis. Furthermore, YTHDF2 overexpression exacerbated the decrease in cell viability under BkF exposure, while YTHDF2 knockdown was the opposite. The results from the RIP assay demonstrated a significant enhancement in the interaction of YTHDF2 protein with BCL2 mRNA following the overexpression of YTHDF2. In addition, animal experiments showed that there was an increase in apoptosis and a decrease in proliferation of testicular cells in mice in the high-dose (30 mg/kg) BkF group by TUNEL staining and Ki67 staining. Immunohistochemical analysis showed that BCL2 levels were significantly lower in the high-dose group than in the control group, while YTHDF2, P53 and BAX were dramatically increased. In summary, our study suggests that YTHDF2 has been implicated in BkF-induced male reproductive injury by promoting the degradation of BCL2.

中文翻译：

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RNA 读取蛋白 YTHDF2 通过调节 BCL2 的稳定性介导苯并（k）荧蒽诱导的男性生殖损伤


苯并 （k） 荧蒽 （BkF） 对雄性生殖有不利影响，但其具体作用机制仍不清楚。本研究重点研究了 RNA 读取蛋白 YTHDF2 的作用及其在 BkF 诱导的男性生殖损伤中的作用机制。小鼠 GC-2 精母细胞暴露于 0 、 40 、 80 、 160 μM BkF 中。研究发现，BkF 显著增加 GC-2 细胞凋亡，降低其存活率。精母细胞中 BCL2 显著降低，而 P53 和 BAX 的表达显著增加。有趣的是，RNA 读取蛋白 YTHDF2 的表达随着 BkF 暴露剂量的增加而逐渐升高。YTHDF2 过表达显著降低细胞活力，增加细胞凋亡率。同时，P53 和 BAX 的表达显著增加，BCL2 显著下调。相反，干扰 YTHDF2 会增加细胞增殖并减少细胞凋亡。此外，YTHDF2 过表达加剧了 BkF 暴露下细胞活力的降低，而 YTHDF2 敲低则相反。RIP 检测结果表明，过表达 YTHDF2 后 YTHDF2 蛋白与 BCL2 mRNA 的相互作用显著增强。此外，动物实验显示，通过 TUNEL 染色和 Ki67 染色，高剂量 （30 mg/kg） BkF 组小鼠细胞凋亡增加，睾丸细胞增殖减少。免疫组化分析显示，高剂量组 BCL2 水平显著低于对照组，而 YTHDF2 、 P53 和 BAX 显著升高。 总之，我们的研究表明，YTHDF2 通过促进 BCL2 的降解与 BkF 诱导的男性生殖损伤有关。